DCL2 | Dicer-like protein 2
AS15 3100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 5000-1 : 10 000 (WB)|
|Expected | apparent MW||
156.9 | 157 kDa
|Confirmed reactivity||Arabidopsis thaliana|
|Not reactive in||
Nicotiana tabacum, Zea mays
to be added when available, antibody released in January 2016.
50 µg of total protein from Arabidopsis thaliana whole vegetative rosette, DCL2 overexpression line (a), wild type Col-0 (b), dcl2-1 intron insertion (c), extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with laemmli buffer at 95°C/5 min. were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Amersham ECL Prime and expose to Amersham Hyperfilms ECL for 120 seconds.
Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
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